Biodegradation of low-density polyethylene by mixed fungi composed of Alternaria sp. and Trametes sp. isolated from landfill sites.

With the development of industry and modern manufacturing, nondegradable low-density polyethylene (LDPE) has been widely used, posing a rising environmental hazard to natural ecosystems and public health. In this study, we isolated a series of LDPE-degrading fungi from landfill sites and carried out LDPE degradation experiments by combining highly efficient degrading fungi in pairs. The results showed that the mixed microorganisms composed of Alternaria sp. CPEF-1 and Trametes sp. PE2F-4 (H-3 group) had a greater degradation effect on heat-treated LDPE (T-LDPE). After 30 days of inoculation with combination strain H-3, the weight loss rate of the T-LDPE film was approximately 154% higher than that of the untreated LDPE (U-LDPE) film, and the weight loss rate reached 0.66 ± 0.06%. Environmental scanning electron microscopy (ESEM) and Fourier transform infrared spectroscopy (FTIR) were used to further investigate the biodegradation impacts of T-LDPE, including the changes on the surface and depolymerization of the LDPE films during the fungal degradation process. Our findings revealed that the combined fungal treatment is more effective at degrading T-LDPE than the single strain treatment, and it is expected that properly altering the composition of the microbial community can help lessen the detrimental impact of plastics on the environment.

Polysaccharides from Trametes versicolor as a Potential Prebiotic to Improve the Gut Microbiota in High-Fat Diet Mice.

Polysaccharides derived from Trametes versicolor have been found to exhibit hypolipidemic activity in hyperlipidemic mice, but the mechanism by which they modulate intestinal flora is still unclear. Currently, this study aimed to investigate the regulatory effects of extracellular (EPTV) and intracellular polysaccharides from T. versicolor (IPTV) on the dysbiosis of intestinal flora in mice fed a high-fat diet (HFD). The results showed that the oral administration of T. versicolor polysaccharides significantly ameliorated lipid accumulation and steatosis in hepatocytes. The gut dysbiosis in the HFD mice was characterized by a decrease in abundance and diversity of bacteria and an increase in the Firmicutes/Bacteroidetes ratio. However, T. versicolor polysaccharides attenuated these changes and reduced the relative abundance of bile-salt-hydrolase (BSH)-producing bacteria, such as Bacillus, Enterococcus, Bifidobacterium, and Lactococcus. It is noteworthy that T. versicolor polysaccharides also restored the disorganization of intestinal fungi in HFD mice, with EPTV treatment leading to a higher relative abundance of Basidiomycota and Ascomycota compared to IPTV. Additionally, T. versicolor polysaccharides enhanced the growth of butyrate-producing bacteria via the buk and but pathways, accompanied by an increase in short-chain fatty acids (SCFAs), especially butyrate. IPTV also increased the expression of G-protein-coupled receptors 41 (GPR41) and 43 (GPR43) by 40.52% and 113.24% each, as compared to 62.42% and 110.28%, respectively, for EPTV. It is suggested that IPTV and EPTV have the potential to counteract hyperlipidemia-associated intestinal flora disorders and improve lipid metabolism.

Macrofungal Extracts as a Source of Bioactive Compounds for Cosmetical Anti-Aging Therapy: A Comprehensive Review.

For centuries, mushrooms have been used as a component of skincare formulations. Environmental stresses and a modern lifestyle expose the skin to accelerated aging. To slow down this process, natural anti-aging skincare ingredients are being sought. In this review, 52 scientific publications about the effects of chemical compounds extracted from the fruiting bodies of macrofungi on skin cells were selected. The effects of extracts from nine species that are tested for anti-aging effects have been described. According to available literature data, macrofungi contain many polysaccharides, phenolic compounds, polysaccharide peptides, free amino acids, sterols, proteins, glycosides, triterpenes, alkaloids, which can have an anti-aging effect on the skin by acting as antioxidants, photoprotective, skin whitening, moisturizing, anti-inflammatory and stabilizing collagen, elastin and hyaluronic acid levels in the skin.

Safety evaluation of an extension of use of the food enzyme laccase from the non-genetically modified Trametes hirsuta strain AE-OR.

The food enzyme laccase (benzenediol:oxygen oxidoreductase, i.e. EC 1.10.3.2) is produced with the non-genetically modified Trametes hirsuta strain AE-OR by Amano Enzyme Inc. A safety evaluation of this food enzyme was made previously, in which EFSA concluded that this food enzyme did not give rise to safety concerns when used in six food manufacturing processes. Subsequently, the applicant has requested to extend its use to include three additional processes and to revise the use levels. In this assessment, EFSA updated the safety evaluation of this food enzyme when used in a total of nine food manufacturing processes. Dietary exposure to the food enzyme-total organic solids (TOS) was calculated to be up to 0.030 mg TOS/kg body weight (bw) per day in European populations. Using the no observed adverse effect level previously reported (862 mg TOS/kg bw per day, the highest dose tested), the Panel derived a margin of exposure of at least 28,733. Based on the data provided for the previous evaluation and the revised margin of exposure in the present evaluation, the Panel concluded that this food enzyme does not give rise to safety concerns under the revised intended conditions of use.

Effects of Fungal Solid-State Fermentation on the Profile of Phenolic Compounds and on the Nutritional Properties of Grape Pomace.

Grape pomace (GP) is considered a natural source of bioactive compounds. To improve the extractability of bioactive compounds, in this work, GP was biologically treated for 15 days with the white-rot fungus Trametes versicolor in laboratory jars and a tray bioreactor under solid-state fermentation (SSF) conditions. During SSF, the activity of lignolytic (laccase and manganese peroxidase) and hydrolytic (xylanase, cellulase, ß-glucosidase, and invertase) enzymes was measured, with the activities of laccase (2.66 U/gdb in jars and 0.96 U/gdb in the bioreactor) and xylanase (346.04 U/gdb in jars and 200.65 U/gdb in the bioreactor) being the highest. The effect of the complex enzyme system was reflected in the changes in the chemical composition of GP with increasing ash, crude protein, and free fat content: 28%, 10%, and 17% in the laboratory jars, and 29%, 11%, and 7% in the bioreactor, respectively. In addition, the biological treatment improved the extractability of 13 individual phenolic compounds. Therefore, the applied SSF technique represents an effective strategy to improve the profile of phenolic compounds and the nutritional composition of GP, promoting their valorization and opening the door for potential applications in the food industry and other sectors.

Structure and Bioactivity of Intracellular and Extracellular Polysaccharides of Trametes lactinea Mycelium.

Trametes lactinea polysaccharides have a high medicinal value; however, we still know little about the structure and bioactivity of intracellular and extracellular polysaccharides in the mycelial liquid fermentation of T. lactinea. This study analyzed the structures of intracellular (IP-1, IP-2, and IP-3) and extracellular (EP-1 and EP-2) polysaccharide components isolated from T. lactinea liquid fermentation, as well as investigated their antioxidant, antibacterial, and immunomodulatory properties. The results showed that IP-3 was the only component with a triple-helix structure, while the other four components did not possess this structure. IP3 has a higher molecular weight, flavonoid, and total phenolic content compared to other components. Both intracellular and extracellular polysaccharide components exhibited strong scavenging abilities against ABTS and DPPH radicals. The components showed limited antibacterial effects against four types of bacteria (Staphylococcus aureus, Bacillus subtilis, Erwinia carotovora, and Escherichia coli), and were found to be non-toxic to RAW264.7 cells, even promoting cell proliferation. Furthermore, within a specific concentration range, all components enhanced the phagocytic activity of RAW264.7 cells, increased the secretion of NO, TNF-α, and IL-6, and demonstrated concentration-dependent effects, with IP-3 displaying the most potent immunomodulatory activity. This study shows a high potential for the development and utilization of polysaccharides derived from the liquid fermentation of T. lactinea mycelium.

Submerged cultivation and phytochemical analysis of medicinal mushrooms (Trametes sp.).

Mushrooms are widely available around the world and have various nutritional as well as therapeutic values. Many Asian cultures believe that medicinal mushrooms can prolong life and improve vitality. This study aims to characterize the phytochemical and polysaccharide content, mainly ß-glucan content, of mycelial biomass and fruiting bodies collected from the Himalayan region, particularly Uttarakhand. Through molecular analysis of the LSU F/R-rDNA fragment sequence and phylogenetic analysis, the strain was identified as Trametes sp. We performed screening of phytochemicals and polysaccharides in mushroom and biomass extracts using high-performance liquid chromatography (HPLC) and a PC-based UV-Vis spectrophotometer. The macrofungal biomass was found to be high in saponin, anthraquinone, total phenolic, flavonoid, and ß-glucan content. In biomass extract, we observed a high level of saponin (70.6µg/mL), anthraquinone (14.5µg/mL), total phenolic (12.45 µg/mL), and flavonoid (9.500 µg/mL) content. Furthermore, we examined the contents of alkaloids, tannins, terpenoids, and sterols in the biomass and mushroom extracts; the concentration of these compounds in the ethanol extract tested was minimal. We also looked for antioxidant activity, which is determined in terms of the IC50 value. Trametes sp. mushroom extract exhibits higher DPPH radical scavenging activity (62.9% at 0.5 mg/mL) than biomass extract (59.19% at 0.5 mg/mL). We also analyzed ß-glucan in Trametes sp. from both mushroom and biomass extracts. The biomass extract showed a higher ß-glucan content of 1.713 mg/mL than the mushroom extract, which is 1.671 mg/mL. Furthermore, ß-glucan analysis was confirmed by the Megazyme ß-glucan assay kit from both biomass and mushroom extract of Trametes sp. ß-glucans have a promising future in cancer treatment as adjuncts to conventional medicines. Producing pure ß-glucans for the market is challenging because 90-95% of ß glucan sold nowadays is thought to be manipulated or counterfeit. The present study supports the recommendation of Trametes sp. as rich in ß-glucan, protein, phytochemicals, and antioxidant activities that help individuals with cancer, diabetes, obesity, etc.

Effects of Orange Peel Extract on Laccase Activity and Gene Expression in Trametes versicolor.

The genome of Trametes versicolor encodes multiple laccase isozymes, the expression of which is responsive to various conditions. Here, we set out to investigate the potential of orange peel extract as an inducer of laccase production in this white-rot fungus, in comparison to the previously identified inducing chemical compound, veratryl alcohol. For four geographically distinct T. versicolor strains, a positive correlation has been observed between their oxidative activity and incubation time in liquid cultures. The addition of 20% orange peel extract or 5 mM veratryl alcohol caused a rapid increase in the oxidative potential of T. versicolor M99 after 24 h, with a more pronounced effect observed for the orange peel extract. To elucidate the underlying molecular mechanisms of the induced laccase activity, a transcriptional gene expression analysis was performed for the seven individual laccase genes in T. versicolor, revealing the upregulation of several laccase genes in response to the addition of each inducer. Notably, the gene encoding TvLac5 demonstrated a substantial upregulation in response to the addition of 20% orange peel extract, likely contributing to the observed increase in its oxidative potential. In conclusion, our results demonstrate that orange peels are a promising agro-industrial side stream for implementation as inducing agents in large-scale laccase production with T. versicolor.

Hydrothermal liquefaction aqueous phase mycoremediation to increase inorganic nitrogen availability.

Hydrothermal liquefaction aqueous phase (HTL-AP) is a waste product from a thermochemical process where wet biomass is converted into biocrude oil. This nutrient-rich wastewater may be repurposed to benefit society by assisting crop growth after adequate treatment to increase inorganic nitrogen, especially NO3 -. This study aims to increase HTL-AP inorganic nitrogen, specifically NH3/NH4 + and NO3 -, through fungal remediation for further use in hydroponic systems. Trametes versicolor, a white-rot fungus known for degrading a range of organic pollutants, was used to treat a diluted (5 %) HTL-AP for 9 days. No fungal growth was observed, but T. versicolor activity was suspected by laccase activity throughout cultivation time. NO3 --N and NH3/NH4 +-N increased by 17 and 8 times after three days of fungal treatment, which was chosen as the appropriate time for HTL-AP fungal treatment as it resulted in the highest concentration of NO3 --N. The addition of nitrifying bacteria to the fungal treatment resulted in a twofold increase in NO3 --N concentration compared to the fungal treatment alone, indicating an enhancement in treatment efficacy. COD decreased by 51.33 % after 24 h, which may be related to the fungus' capacity to reduce the concentration of organics in the wastewater; nonetheless, COD increased in the following days, which may be related to the release of fungal byproducts. T. versicolor shows promise as a potential candidate for increasing inorganic nitrogen in HTL-AP. However, future studies should primarily address HTL-AP toxicity, reducing NH3/NH4 +-N while increasing NO3 --N, and hydroponics crop production after fungal treatment.

Toxicological study on ibuprofen and selenium in freshwater mussel Lamellidens marginalis and exploring the microbial cytochrome through modelling and quantum mechanics approaches for its toxicity degradation in contaminated environment.

Toxicological stress in aquatic organisms is caused by the discharge of hundreds of toxic pollutants and contaminants among which the current study concentrates on the toxic effect of non-steroidal anti-inflammatory drug ibuprofen (IBF) and the trace element selenium (Se). In this study, IBF and Se toxicity on freshwater mussel Lamellidens marginalis was studied for 14 days, and in silico predictions for their degradation were made using Molecular modelling and Quantum Mechanical approaches. The degrading propensity of cytochrome c oxidase proteins from Trametes verticillatus and Thauera selenatis (Turkey tail fungi and Gram-negative bacteria) is examined into atom level. The results of molecular modelling study indicate that ionic interactions occur in the T. selenatis-HEME bound complex by Se interacting directly with HEME, and in the T. versicolor-HEME bound complex by IBF bound to a nearby region of HEME. Experimental and theoretical findings suggest that, the toxicological effects of Se and IBF pollution can be reduced by bioremediation with special emphasis on T. versicolor, and T. selenatis, which can effectively interact with Se and IBF present in the environment and degrade them. Besides, this is the first time in freshwater mussel L. marginalis that ibuprofen and selenium toxicity have been studied utilizing both experimental and computational methodologies for their bioremediation study.

Isolation, Identification, Phytochemical Characterization, and In Vitro Anti-Inflammatory Property of an Endophytic Trametes versicolor CL-1 Isolated from Rosa roxburghii.

This study explores the anti-inflammatory potential of an endophytic fungus, Trametes versicolor CL-1, isolated from the fruit tissues of Rosa roxburghii. Morphological and molecular analyses confirmed the identity of CL-1. An ethyl acetate extract (CL-E) from its fermentation broth was subjected to UPLC-HRMS and GNPS molecular networking. The analysis revealed a diverse array of secondary metabolites, including 11 terpenes, 7 flavonoids, 10 cinnamic acid derivatives, 6 oligopeptides, and 9 fatty acids, as verified by LC-MS/MS. Notably, CL-E exhibited significant in vitro anti-inflammatory activity in RAW264.7 cells. Furthermore,  molecular docking studies predicted favorable binding interactions of key compounds 1 within CL-E with the NLRP3 inflammasome (PDB ID: 6NPY). These findings suggest T. versicolor CL-1 as a promising source of natural anti-inflammatory agents and unveil R. roxburghii as a potential reservoir for discovering novel bioactive metabolites.

Biofoams with untapped enzymatic potential produced from beer bagasse by indigenous fungal strains.

White rot fungi are promising organisms for the production of mycelial-based biofoams, providing a sustainable means of valorizing lignocellulosic wastes. This study explores the utilization of two indigenous fungal species, isolated from Argentina and belonging to the genera Trametes, for producing biofoams from brewery waste. The resulting biofoams exhibited an average density of 0.30 g cm-3, a Young's modulus of approximately 1 MPa, and a compressive stress of around 19 MPa. Additionally, the variation of laccase activity throughout the biofoam production process was evaluated. Surprisingly, residual laccase activity was detected in the biofoams following oven drying at temperatures of 60, 80, and 100 °C. This detection highlights the untapped enzymatic potential of the biofoams and positions them as promising green catalysts for various biotechnological applications.

Development of binderless fiberboard from poplar wood residue with Trametes hirsuta.

The utilization of agricultural and forestry residues for the development and preparation of green binderless fiberboard (BF) is an effective way to realize high-value utilization of lignocellulose biomass resources. This study focuses on the fabrication of BF with excellent mechanical and waterproof properties, utilizing poplar wood residue (PWR) as raw material and Trametes hirsuta as a pretreatment method. During the fermentation process, lignin-degrading enzymes and biological factors, such as sugars, were produced by T. hirsuta, which activated lignin by depolymerizing lignin bonds and modifying structural functional groups, and forming new covalent bonds between poplar fibers, ultimately enhancing adhesion. Additionally, the activated lignin molecules and sugar molecules coalesce under high temperatures and pressures, forming a dense carbonization layer that bolsters the mechanical properties of the fiberboard and effectively shields it from rapid water infiltration. The bio-pretreated BF for 10 days shows a MOR and MOE of up to 36.1 Mpa and 3704.3 Mpa, respectively, which is 261% and 247.8% higher than that of the bio-untreated fiberboard, and the water swelling ratio (WSR) rate is only 5.6%. Chemical composition analysis revealed that repolymerization occurred among lignin, cellulose, and hemicellulose, especially the molecular weight of lignin changed significantly, with the Mw of lignin increasing from 312066 g/mol to 892362 g/mol, and then decreasing to 825021 g/mol. Mn increased from 277790 g/mol to 316987.5 g/mol and then decreased to 283299.5 g/mol at 21 days. Compared to other artificial fiberboards prepared through microbial pretreatment, the BF prepared by microorganisms in this study exhibited the highest mechanical properties among the poplar wood biobased panels.

Long-term oral administration of Huaier granules improves survival outcomes in hepatocellular carcinoma patients within Milan criteria following microwave ablation: a propensity score matching and stabilized inverse probability weighting analysis.

Objective: This study aimed to elucidate the therapeutic effects of Huaier granules on hepatocellular carcinoma (HCC) within the Milan criteria in patients who underwent microwave ablation (MWA). Materials and methods: A total of 228 patients were included, with 97 in the Huaier group and 131 in the control group. We evaluated progression-free survival (PFS), overall survival (OS), and extrahepatic metastasis survival (EMS) using Kaplan-Meier (KM) curves with a log-rank test. Propensity Score Matching (PSM) and Stabilized Inverse Probability of Treatment Weighting (IPTW) were performed to minimize selection and confounding biases. Results: Following PSM, the 1-, 3-, and 5-year PFS rates in the Huaier and control groups were 83.5% vs 70.7%, 57.7% vs 42.6%, and 43.6% vs 31.9% (p = 0.030), respectively. The 1-, 3-, and 5-year OS rates were 98.9% vs 95.6%, 83.9% vs 72.3%, and 72.2% vs 53.7% (p = 0.023), respectively. The corresponding 1-, 3-, and 5-year EMS rates were 98.9% vs 93.4%, 91.7% vs 83.7%, and 91.7% vs 78.5% (p = 0.039), respectively. Stabilized IPTW analysis of KM curves yielded results similar to those of the PSM analysis. Additionally, administering Huaier granules for at least 6 months significantly improved PFS and OS. Conclusion: Huaier granules can reduce the risk of recurrence and improve the OS of patients with HCC within the Milan criteria following MWA. Administering Huaier granules for over 6 months proved beneficial.

Effects of Environmental and Nutritional Conditions on Mycelium Growth of Three Basidiomycota.

In recent decades, an enormous potential of fungal-based products with characteristics equal to, or even outperforming, classic petroleum-derived products has been acknowledged. The production of these new materials uses mycelium, a root-like structure of fungi consisting of a mass of branching, thread-like hyphae. Optimizing the production of mycelium-based materials and fungal growth under technical conditions needs to be further investigated. The main objective of this study was to select fast-growing fungi and identify optimized incubation conditions to obtain a dense mycelium mat in a short time. Further, the influence of the initial substrate characteristics on hyphae expansion was determined. Fungal isolates of Ganoderma lucidum, Pleurotus ostreatus, and Trametes versicolor were cultivated for seven days on substrate mixtures consisting of various proportions of pine bark and cotton fibers. Furthermore, the substrates were mixed with 0, 2, and 5 wt.% calcium carbonate (CaCO3), and the incubator was flushed with 0, 5, and 10 vol.% carbon dioxide (CO2). All samples grew in the dark at 26 °C and a relative humidity of 80%. Evaluation of growth rate shows that cotton fiber-rich substrates performed best for all investigated fungi. Although Pleurotus ostreatus and Trametes versicolor showed comparatively high growth rates of up to 5.4 and 5.3 mm d-1, respectively, mycelium density was thin and transparent. Ganoderma lucidum showed a significantly denser mycelium at a maximum growth rate of 3.3 mm d-1 on a cotton fiber-rich substrate (75 wt.%) without CaCO3 but flushed with 5 vol.% CO2 during incubation.

Safety evaluation of the food enzyme laccase from the non-genetically modified Trametes hirsuta strain AE-OR.

The food enzyme laccase (benzenediol:oxygen oxidoreductase; EC 1.10.3.2) is produced with the non-genetically modified Trametes hirsuta strain AE-OR by Amano Enzyme Inc. The food enzyme is free from viable cells of the production organism. It is intended to be used in six food manufacturing processes. Dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.026 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 862 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 33,154. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that a risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Trametes versicolor Laccase-Based Magnetic Inorganic-Protein Hybrid Nanobiocatalyst for Efficient Decolorization of Dyes in the Presence of Inhibitors.

In the present investigation, an ecofriendly magnetic inorganic-protein hybrid system-based enzyme immobilization was developed using partially purified laccase from Trametes versicolor (TvLac), Fe3O4 nanoparticles, and manganese (Mn), and was successfully applied for synthetic dye decolorization in the presence of enzyme inhibitors. After the partial purification of crude TvLac, the specific enzyme activity reached 212 U∙mg total protein-1. The synthesized Fe3O4/Mn3(PO4)2-laccase (Fe3O4/Mn-TvLac) and Mn3(PO4)2-laccase (Mn-TvLac) nanoflowers (NFs) exhibited encapsulation yields of 85.5% and 90.3%, respectively, with relative activities of 245% and 260%, respectively, compared with those of free TvLac. One-pot synthesized Fe3O4/Mn-TvLac exhibited significant improvements in catalytic properties and stability compared to those of the free enzyme. Fe3O4/Mn-TvLac retained a significantly higher residual activity of 96.8% over that of Mn-TvLac (47.1%) after 10 reuse cycles. The NFs showed potential for the efficient decolorization of synthetic dyes in the presence of enzyme inhibitors. For up to five reuse cycles, Fe3O4/Mn-TvLac retained a decolorization potential of 81.1% and 86.3% for Coomassie Brilliant Blue R-250 and xylene cyanol, respectively. The synthesized Fe3O4/Mn-TvLac showed a lower acute toxicity towards Vibrio fischeri than pure Fe3O4 nanoparticles did. This is the first report of the one-pot synthesis of biofriendly magnetic protein-inorganic hybrids using partially purified TvLac and Mn.

Biodegradation of Trichloroethylene by Trametes versicolor and its Physiological Response to Contaminant Stress.

Trichloroethylene (TCE) poses a potentially toxic threat to humans and the environment and widely exists in contaminated sites. White rot fungi effectively degrade refractory pollutants, while a few research studies use white rot fungi to degrade TCE. In this study, we investigated TCE biodegradation by white rot fungi and the potential influencing factors in the environment and attempted to research the effect of TCE on the physiological characteristics of white rot fungi. White rot fungi (Trametes versicolor, Pseudotrametes gibbosa, Pycnoporus sanguines and Pleurotus ostreatus) were added to the liquid medium for shock culture. The results revealed that T. versicolor exhibited the most pronounced efficacy in removing TCE, with a degradation rate of 81.10% within a 7 d period. TCE induces and is degraded by cytochrome P450 enzymes. High pH and Cr(VI) adversely affected the effectiveness of the biodegradation of TCE, but the salinity range of 0-1% had less effect on biodegradation. Overall, the effectiveness of degradation of TCE by T. versicolor has been demonstrated, and it provides a reference for the application prospects of white rot fungi in TCE-contaminated soils.

Structure elucidation and antiviral activity of a cold water-extracted mannogalactofucan Ts1-1A from Trametes sanguinea against human cytomegalovirus in vitro.

In this study, we purified a partially acetylated heteropolysaccharide (Ts1-1A) from the fruit bodies of Trametes sanguinea Lloyd through cold water extraction and serial chromatographic separation. The purified polysaccharide Ts1-1A (12.8 kDa) was characterized as a branched mannogalactofucan with a backbone of alternately connected 1,3-linked α-Fucp and 1,6-linked α-Galp, which was partially substituted by non-reducing end units of ß-Manp at O-2 and O-3 positions of 1,6-linked α-Galp. Ts1-1A showed pronounced anti-human cytomegalovirus activity at the concentration of 200 and 500 µg/mL in systematical assessments including morphological changes, western blotting, qPCR, indirect immunofluorescence and tissue culture infective dose assays. Moreover, Ts1-1A exerted its antiviral activity at two distinct stages of viral proliferation manifesting as significantly inhibiting viral protein (IE1/2 and p52) expression and reducing viral gene (UL123, UL44 and UL32) replication in the HCMV-infected WI-38 cells. At viral attachment stage, Ts1-1A interacted with HCMV and prevented HCMV from attaching to its host cells. While at early phase of viral replication stage, Ts1-1A suppressed HCMV replication by downregulating NQO1 and HO-1 proteins related to oxidative stress as an antioxidant. To sum up, Ts1-1A is a promising anti-HCMV agent which could be developed for HCMV infection prevention and therapy.

Use of sawdust for production of ligninolytic enzymes by white-rot fungi and pharmaceutical removal.

Use of white-rot fungi for enzyme-based bioremediation of wastewater is of high interest. These fungi produce considerable amounts of extracellular ligninolytic enzymes during solid-state fermentation on lignocellulosic materials such as straw and sawdust. We used pure sawdust colonized by Pleurotus ostreatus, Trametes versicolor, and Ganoderma lucidum for extraction of ligninolytic enzymes in aqueous suspension. Crude enzyme suspensions of the three fungi, with laccase activity range 12-43 U/L and manganese peroxidase activity range 5-55 U/L, were evaluated for degradation of 11 selected pharmaceuticals spiked at environmentally relevant concentrations. Sulfamethoxazole was removed significantly in all treatments. The crude enzyme suspension from P. ostreatus achieved degradation of wider range of pharmaceuticals when the enzyme activity was increased. Brief homogenization of the colonized sawdust was also observed to be favorable, resulting in significant reductions after a short exposure of 5 min. The highest reduction was observed for sulfamethoxazole which was reduced by 84% compared to an autoclaved control without enzyme activity and for trimethoprim which was reduced by 60%. The compounds metoprolol, lidocaine, and venlafaxine were reduced by approximately 30% compared to the control. Overall, this study confirmed the potential of low-cost lignocellulosic material as a substrate for production of enzymes from white-rot fungi. However, monitoring over time in bioreactors revealed a rapid decrease in enzymatic ligninolytic activity.